WO1988010308A1 - Promoteur de levure induisible par pheromones - Google Patents

Promoteur de levure induisible par pheromones Download PDF

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Publication number
WO1988010308A1
WO1988010308A1 PCT/US1988/002129 US8802129W WO8810308A1 WO 1988010308 A1 WO1988010308 A1 WO 1988010308A1 US 8802129 W US8802129 W US 8802129W WO 8810308 A1 WO8810308 A1 WO 8810308A1
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WO
WIPO (PCT)
Prior art keywords
dna
protein
polypeptide
yeast
promoter
Prior art date
Application number
PCT/US1988/002129
Other languages
English (en)
Inventor
Gerald R. Fink
Joshua Trueheart
Elaine A. Elion
Original Assignee
Whitehead Institute For Biomedical Research
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Whitehead Institute For Biomedical Research filed Critical Whitehead Institute For Biomedical Research
Publication of WO1988010308A1 publication Critical patent/WO1988010308A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2468—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1) acting on beta-galactose-glycoside bonds, e.g. carrageenases (3.2.1.83; 3.2.1.157); beta-agarase (3.2.1.81)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/37—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi
    • C07K14/39—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi from yeasts
    • C07K14/395—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi from yeasts from Saccharomyces
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405—Glucanases
    • C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
    • C12N9/2431—Beta-fructofuranosidase (3.2.1.26), i.e. invertase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01023—Beta-galactosidase (3.2.1.23), i.e. exo-(1-->4)-beta-D-galactanase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01026—Beta-fructofuranosidase (3.2.1.26), i.e. invertase
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/61—Fusion polypeptide containing an enzyme fusion for detection (lacZ, luciferase)

Definitions

  • the resulting plasmid is introduced into wild-type a_ cells, which are incubated with alpha factor.
  • Alpha factor induces a-type cells to respond; induction of the FUSl promoter leads to induction of transcription of the DNA of interest to which it is fused.
  • the result is expression of the FUSl-DNA of interest fusion protein (i.e., a fusion protein which is the FUSl-encoded protein and the protein encoded by the DNA of interest) at substantial levels.
  • yeast S. cerevisiae can exist in a diploid or a haploid state; in the former, it has 17 pairs of chromosomes and, in the latter, a single set of chromosomes (one copy of each of the 17 chromosomes) . Under favorable conditions, yeast cells exist in the diploid state and multiply rapidly. Multiplication occurs through mitosis, the process by which a diploid cell duplicates its chromosome pairs and divides into a mother cell and a bud, each of which has two copies of every chromosome.
  • Conjugation in S. cerevisiae involves the fusion of haploid cells of opposite mating type, followed by the fusion of nuclei to form a diploid.
  • the zygote formed by this process buds off diploid cells capable of vegetative growth. Formation of the zygote requires the coordination of two processes—cell fusion and nuclear fusion. Both processes are initiated by mating phero ones: a. cells produce a.-factor, to which alpha cells specifically respond, and alpha cells produce alpha-factor, to which ⁇ cells specifically respond. Duntze, W.D. et al. , European Journal of Biochemistry, 3_5:357-365 (1973); Wilkinson, L.E. and J.R. Pringle, Experimental Cell Research, 8_9:175-187 (1974).
  • a recombinant plasmid which can be used in the method of the present invention, the following components are included: a) a fragment of a plasmid, such as pBR322, which includes the plasmid DNA replication origin, which makes it possible to propagate
  • the fus2-l mutation was uncovered in a cross between C52a and JY145 (fusl-483) .
  • Several tetrads from this cross produced one ascospore which, when mated to ⁇ 453 lawns, displayed a drastic defect as compared to the response with wild type lawns. (fusl alone displays only a partial defect when mated to ⁇ 453).

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Mycology (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Biophysics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

Un promoteur de levure est induit par une phéromone appropriée et un procédé d'expression d'un gène voulu en quantités considérables consiste à la placer sous le contrôle du promoteur induisible. De l'ADN de codage d'une protéine voulue est fusionné ou relié à un promoteur de levure induisible par une phéromone, tel que les promoteurs FUS1 ou FUS2, et leur fusion est insérée dans un vecteur permettant de produire un nombre élevé de copies; le produit ainsi obtenu est introduit dans des cellules de levure de type sauvage. La stimulation de ces cellules de levure par la phéromone appropriée entraîne l'induction de la transcriptase du promoteur de levure et l'expression de quantités considérables de la protéine voulue.
PCT/US1988/002129 1987-06-24 1988-06-23 Promoteur de levure induisible par pheromones WO1988010308A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US6607887A 1987-06-24 1987-06-24
US066,078 1987-06-24

Publications (1)

Publication Number Publication Date
WO1988010308A1 true WO1988010308A1 (fr) 1988-12-29

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Application Number Title Priority Date Filing Date
PCT/US1988/002129 WO1988010308A1 (fr) 1987-06-24 1988-06-23 Promoteur de levure induisible par pheromones

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WO (1) WO1988010308A1 (fr)

Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998013513A2 (fr) * 1996-09-24 1998-04-02 Cadus Pharmaceutical Corporation Procedes et compositions pour identifier des modulateurs de recepteur
EP0915154A1 (fr) * 1993-03-31 1999-05-12 Cadus Pharmaceuticals, Inc. Cellules de levure traítées pour produire des substituts de protéines du système de phéromones, et leurs emplois
WO1999060138A2 (fr) * 1998-05-16 1999-11-25 The Victoria University Of Manchester Expression regulee de pkc et/ou de srb1/psa1 dans la levure
US6251605B1 (en) 1998-10-27 2001-06-26 Cadus Pharmaceutical Corporation Yeast cells having mutations in Cav1 and uses therefor
US6255059B1 (en) 1993-03-31 2001-07-03 Cadus Pharmaceutical Corporation Methods for identifying G protein coupled receptor effectors
US6355473B1 (en) 1998-05-06 2002-03-12 Cadus Pharmaceutical Corp. Yeast cells having mutations in stp22 and uses therefor
US6504008B1 (en) 1999-02-01 2003-01-07 Cadus Technologies, Inc. Cell based signal generation
US6555325B1 (en) 1999-06-14 2003-04-29 Cadus Technologies, Inc. System for detection of a functional interaction between a compound and a cellular signal transduction component
US6864060B1 (en) 1993-03-31 2005-03-08 Cadus Technologies, Inc. Yeast cells expressing modified G proteins and methods of use therefor
US7081360B2 (en) 1998-07-28 2006-07-25 Cadus Technologies, Inc. Expression of G protein-coupled receptors with altered ligand binding and/or coupling properties
US7105309B2 (en) 1993-03-31 2006-09-12 Cadus Technologies, Inc. Yeast cells engineered to produce pheromone system protein surrogates and uses therefor
US7223533B2 (en) 1999-09-10 2007-05-29 Cadus Technologies, Inc. Cell surface proteins and use thereof as indicators of activation of cellular signal transduction pathways
US7235648B1 (en) 1993-03-31 2007-06-26 Cadus Technologies, Inc. Yeast cells engineered to produce pheromone system protein surrogates, and uses therefor
US7250263B2 (en) 1993-03-31 2007-07-31 Cadus Technologies, Inc. Methods and compositions for identifying receptor effectors
US7273747B2 (en) 1998-08-27 2007-09-25 Cadus Technologies, Inc. Cell having amplified signal transduction pathway responses and uses therefor
US7319009B2 (en) 1998-10-07 2008-01-15 Cadus Technologies, Inc. Methods and compositions for identifying receptor effectors
US7416881B1 (en) 1993-03-31 2008-08-26 Cadus Technologies, Inc. Yeast cells engineered to produce pheromone system protein surrogates, and uses therefor

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0123544A2 (fr) * 1983-04-25 1984-10-31 Genentech, Inc. Procédé d'expression de protéines hétérologues dans les levures, vecteurs d'expression et micro-organismes du genre levure à cet effet
EP0183350A2 (fr) * 1984-10-29 1986-06-04 Immunex Corporation ADN codant pour le facteur stimulant des colonies, peptide codé par celui-ci, vecteurs et hôtes transformés contenant un tel ADN et leur préparation
US4615974A (en) * 1981-08-25 1986-10-07 Celltech Limited Yeast expression vectors

Patent Citations (3)

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Publication number Priority date Publication date Assignee Title
US4615974A (en) * 1981-08-25 1986-10-07 Celltech Limited Yeast expression vectors
EP0123544A2 (fr) * 1983-04-25 1984-10-31 Genentech, Inc. Procédé d'expression de protéines hétérologues dans les levures, vecteurs d'expression et micro-organismes du genre levure à cet effet
EP0183350A2 (fr) * 1984-10-29 1986-06-04 Immunex Corporation ADN codant pour le facteur stimulant des colonies, peptide codé par celui-ci, vecteurs et hôtes transformés contenant un tel ADN et leur préparation

Non-Patent Citations (7)

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Title
CHEMICAL ABSTRACTS, Vol. 103, (1985), Abstract No. 208060x, (MIYAJIMA ATUSUSHI); & GENE, 1985, 37(1-3), 155-61 (Eng). *
CHEMICAL ABSTRACTS, Vol. 104, (1986), Abstract No. 16037u, (SINGH ARJUN et al.); & GENET: NEW FRONT., PROC. INT. CONGR., 15th 1983, (Pub. 1984). 2, 33-9 (Eng). *
CHEMICAL ABSTRACTS, Vol. 104, (1986), Abstract No. 220092n, (VLASUK GEORGE P.); & J. BIOL. CHEM., 1986, 261(11), 4789-96 (Eng). *
CHEMICAL ABSTRACTS, Vol. 107, (1987), Abstract No. 128287u, (McCAFFREY); & MOL. CELL. BIOL., 1987, 7(8), 2680-90 (Eng). *
CHEMICAL ABSTRACTS, Vol. 107, (1987), Abstract No. 212743x, (TRUEHEART); & MOL. CELL. BIOL., 1987, 7(7), 2316-28, (Eng). *
CHEMICAL ABSTRACTS, Vol. 108, (1988), Abstract No. 217221f, (NONATO, ROBERTO V.); & BIOCHEM. BIOPHYS. RES. COMMUN., 1988, 152(1), 76-82 (Eng). *
MOLECULAR AND CELLULAR BIOLOGY, Volume 7, No. 2, February 1987, SCOTT W. VAN ARSDELL et al., "The Yeast Repeated Element Sigma Contains a Hormone-Inducible Promoter", pages 749-759. *

Cited By (27)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7250263B2 (en) 1993-03-31 2007-07-31 Cadus Technologies, Inc. Methods and compositions for identifying receptor effectors
US6864060B1 (en) 1993-03-31 2005-03-08 Cadus Technologies, Inc. Yeast cells expressing modified G proteins and methods of use therefor
EP0915154A1 (fr) * 1993-03-31 1999-05-12 Cadus Pharmaceuticals, Inc. Cellules de levure traítées pour produire des substituts de protéines du système de phéromones, et leurs emplois
US7611854B2 (en) 1993-03-31 2009-11-03 Cadus Technologies, Inc. Yeast cells engineered to produce pheromone system protein surrogates, and uses therefor
US7416881B1 (en) 1993-03-31 2008-08-26 Cadus Technologies, Inc. Yeast cells engineered to produce pheromone system protein surrogates, and uses therefor
US7122305B2 (en) 1993-03-31 2006-10-17 Cadus Technologies, Inc. Methods and compositions for identifying receptor effectors
US7105309B2 (en) 1993-03-31 2006-09-12 Cadus Technologies, Inc. Yeast cells engineered to produce pheromone system protein surrogates and uses therefor
US7361498B2 (en) 1993-03-31 2008-04-22 Cadus Technologies, Inc. Yeast cells expressing modified G proteins and methods of use therefor
US6255059B1 (en) 1993-03-31 2001-07-03 Cadus Pharmaceutical Corporation Methods for identifying G protein coupled receptor effectors
US7235648B1 (en) 1993-03-31 2007-06-26 Cadus Technologies, Inc. Yeast cells engineered to produce pheromone system protein surrogates, and uses therefor
WO1998013513A2 (fr) * 1996-09-24 1998-04-02 Cadus Pharmaceutical Corporation Procedes et compositions pour identifier des modulateurs de recepteur
US6159705A (en) * 1996-09-24 2000-12-12 Cadus Pharmaceutical Corporation Recombinant yeast cells for identifying receptor effectors
WO1998013513A3 (fr) * 1996-09-24 1998-09-03 Cadus Pharmaceutical Corp Procedes et compositions pour identifier des modulateurs de recepteur
US6355473B1 (en) 1998-05-06 2002-03-12 Cadus Pharmaceutical Corp. Yeast cells having mutations in stp22 and uses therefor
GB2353798A (en) * 1998-05-16 2001-03-07 Univ Manchester Regulated expression of PKC and/or SRB1/PSA1 in yeast
GB2353798B (en) * 1998-05-16 2003-11-12 Univ Manchester Regulated expression of SRB1/PSA1 in yeast
WO1999060138A3 (fr) * 1998-05-16 2000-01-20 Univ Manchester Expression regulee de pkc et/ou de srb1/psa1 dans la levure
WO1999060138A2 (fr) * 1998-05-16 1999-11-25 The Victoria University Of Manchester Expression regulee de pkc et/ou de srb1/psa1 dans la levure
US7081360B2 (en) 1998-07-28 2006-07-25 Cadus Technologies, Inc. Expression of G protein-coupled receptors with altered ligand binding and/or coupling properties
US7273747B2 (en) 1998-08-27 2007-09-25 Cadus Technologies, Inc. Cell having amplified signal transduction pathway responses and uses therefor
US7319009B2 (en) 1998-10-07 2008-01-15 Cadus Technologies, Inc. Methods and compositions for identifying receptor effectors
US6251605B1 (en) 1998-10-27 2001-06-26 Cadus Pharmaceutical Corporation Yeast cells having mutations in Cav1 and uses therefor
US7022513B2 (en) 1999-02-01 2006-04-04 Cadus Technologies, Inc. Cell based signal generation
US6504008B1 (en) 1999-02-01 2003-01-07 Cadus Technologies, Inc. Cell based signal generation
US7090991B2 (en) 1999-06-14 2006-08-15 Cadus Technologies, Inc. System for detection of a functional interaction between a compound and a cellular signal transduction component
US6555325B1 (en) 1999-06-14 2003-04-29 Cadus Technologies, Inc. System for detection of a functional interaction between a compound and a cellular signal transduction component
US7223533B2 (en) 1999-09-10 2007-05-29 Cadus Technologies, Inc. Cell surface proteins and use thereof as indicators of activation of cellular signal transduction pathways

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